Mendeleev

Communications
Mendeleev Commun., 2022, 32, 136–138

Rapid colorimetric determination of ascorbic acid by solid phase

extraction of iodine into a polymethacrylate matrix
Nadezhda V. Saranchina,a Anna A. Damzina,b Nataliya A. Gavrilenko,b Tatyana N. Volgina,a
Yaroslav E. Ermolaev,a Marina S. Polonskayaa and Mikhail A. Gavrilenko*a
a Tomsk Polytechnic University, 634050 Tomsk, Russian Federation. E-mail: dce@mail.ru
b Tomsk State University, 634050 Tomsk, Russian Federation
DOI: 10.1016/j.mencom.2022.01.044

The proposed iodometric solid-phase spectrophotometry

method is based on interaction of ascorbic acid with
molecular iodine in a solution with further extraction of the
unreacted iodine into a polymethacrylate matrix followed
by its spectrophotometric measurement in the solid phase.
The amount of iodine extracted by the matrix is in inverse
linear dependence on the concentration of ascorbic acid in
the solution. The method can be employed for the range of
ascorbic acid concentrations 1.0–9.0 mg dm–3, while its
detection limit is 0.8 mg dm–3.

Keywords: ascorbic acid, polymethacrylate matrix, solid phase spectrophotometry, iodometric method.

Ascorbic acid (vitamin C, food additive E300) is widely used in
pharmaceutical, perfume, cosmetics and food industries to
increase acidity of products and prolong their shelf life.1,2
Numerous analytical techniques are available for the
determination of ascorbic acid in different matrices. The
instrumental methods such as electrochemical3–8 and
chromatographic ones9–11 are paid a lot of attention. As well, a
significant number of spectrophotometric techniques for the
analysis of ascorbic acid,12–16 including solid-phase ones with
the use of reagents immobilized into a solid support,17–20 have
been developed.
For the officially accepted titrimetric determination of
ascorbic acid,21 2,6-dichloroindophenol {2,6-dichloro-4-
[(4-hydroxyphenyl)imino]-2,5-cyclohexadien-1-one}22–24 is
recommended. As well, ferroin [tris(1,10-phenanthroline)-
iron(ii)]25,26 and Cuii with neocuproine27 are used as reagents
immobilized on solid carriers for the quantitative analysis of
ascorbic acid. The immobilization allows one to carry out the
analysis in a variety of samples including juices with pulp, turbid
and intensely coloured media avoiding preliminarily filtration or
centrifugation to the required transparency. Moreover, this
technique also simplifies the digital image colorimetry for rapid
determination of ascorbic acid in natural fruit juices at the sites
of sample acquisition.28,29
In addition to the above mentioned advantages of the
immobilized reagents, development of a solid-phase analytical
system for direct determination of ascorbic acid skipping the
reagent immobilization stage, i.e., without preliminary
preparation of the solid phase, is of definite interest. This could
significantly simplify the analysis and increase its performance.
Such an approach can be implemented using transformation of
the classical iodometric method of ascorbic acid determination
into a spectrophotometric one to increase the sensitivity.30,31
An example is the analysis of ascorbic acid in food based on its
redox reaction with triiodide ion (I3–) followed by extraction of
the excess I3– into CCl4,32 where iodine forms an associate with
the polymethine dye. In this work, we investigated feasibility
of the thechnique combining solid-phase spectrophotometry
and iodometric method for determination of ascorbic acid
through its oxidation by iodine with further solid-phase
extraction of the unreacted iodine left after the reaction into
polymethacrylate matrix (PMM) and detection of its absorbance
in the solid phase. Solid-phase methods in analytical chemistry
correspond to the green chemistry principles as a key trend in
this area of science.
PMM was used as a transparent analytical medium thanks to
the hydrophilic elements in the PEG 400 chain and the
hydrophobic polymer scaffold of polymethylmethacrylate. The
optical analytical medium had the form of plates obtained by
block polymerization of methacrylic monomers in the presence
of PEG 400 and calcium methacrylate at 60 °C for 3 h initiated
by benzoyl peroxide. The obtained PMM samples represented
transparent colorless plates of 4 × 6 × 0.5 mm size. Combining a
hydrophobic scaffold with a hydrophilic filling to build a
transparent hybrid matrix is a currently relevant and actively
developing approach to promising as ready-to-use analytical
sensors for solid-phase spectrophotometric determinations,
where accumulation of an analyte within the matrix and the
change in the absorption intensity take place through the analyte
extraction from a sample solution.
Determination of ascorbic acid is based on its interaction with
iodine in a solution followed by extraction of the unreacted
iodine into PMM, which results in the change of its color to
yellowish brown in a linear dependence inversely to the
concentration of ascorbic acid in the sample solution. The color
of PMM remains unchanged for a few weeks, with no loss of
analytical response and optical characteristics. Two absorption
maxima at 290 and 365 nm can be seen (Figure 1) in the spectra

© 2022 Mendeleev Communications. Published by ELSEVIER B.V. –  136 –

on behalf of the N. D. Zelinsky Institute of Organic Chemistry of the
Russian Academy of Sciences.
 Mendeleev Commun., 2022, 32, 136–138

2.5

1 1 2 3 4 5 6

Absorbance (arbitrary units)

2.0 1,6
1.4
5 3
1.2 4

A365 (arbitrary units)

1.5 2
1.0
2 1
0.8
1.0
3
0.6
0.4
0.5 4
0.2
5 0.0
0.0 6 0 1 2 3 4 5 6 7 8 9 10
260 310 360 410 460 casc/mg dm–3
l/nm
Figure 1 Absorption spectra of iodine extracted into PMM after its contact Figure 2 Calibration curves for determination of ascorbic acid at different
with a solution containing (1) 0, (2) 1, (3) 3, (4) 5, (5) 7 and (6) HCl concentrations in the sample solution (in 10–3 mol dm–3):
9 mg dm-3 ascorbic acid. Inset: photos of PMM with extracted iodine. (1) 0, (2) 2, (3) 4, (4) 6 and (5) 8. The curve (4) was used further (see the text).

of iodine extracted into PMM after the contact of the matrices influenced the analytical signal with the fixed analyte concentration
2.5 and the iodine one of 5 × 10 mol dm .
with ascorbic acid solutions of various concentrations in the of 5.0 mg dm –3 –5 –3

presence of iodine. We used the absorption maximum of 365 nm Preliminary tests revealed
1
that the analytical signal was recorded
2 3
1 –3 4 5 6
to obtain more repeatable results. at (1.0–8.0) ×2.010–3 mol dm HCl, the corresponding calibration

Absorbance (arbitrary units)

The influence of iodine addition on absorbance at 365 nm was curves A365 vs. casc were collected in Figure 2.
evaluated from calibration curves resulting in their analytical Further experiments
1.5 were carried out with the sample solution
performance as well as the estimated values of regression variance containing 6 × 10–3 mol 2 dm–3 HCl, the corresponding calibration
S2ad and the error mean square S2y (Table 1). Repeatable results curve A365 = 1.423 – 0.161casc (r = 0.998, see Figure 2, curve 4)
1.0
could not be obtained with the iodine concentrations in the sample provided for the widest3 linear response range with an increase in

solution below 5.0 × 10–5 mol dm–3, while the concentrations the sensitivity.0.5 4
³ 1.0 × 10–4 mol dm–3 significantly increased the blank Interfering influence
5 of cations and anions on the
absorbance. We concluded that the optimal iodine concentration determination0.0 of ascorbic
6 acid was evaluated as relative
260 310 360 410 460
in a sample solution for determination of ascorbic acid was deviation of the analytical l/nm signal at casc = 3.0 mg dm–3 and
5.0 × 10–5 mol dm–3, which corresponded to the lowest regression 1 : 1 or 1 : 10 ratios of the analyte to ions. In the presence of
variance and error mean square of a calibration curve as well as NO3¯, NO2¯, Pb2+ or Cu2+ the relative deviation of the analytical
the minimal determined concentration of ascorbic acid. signal exceeded 10%. Addition of EDTA to the sample solution
To test the influence of pH, we explored how 0 to as a masking agent eliminated the influence of the metal
8 × 10–3 mol dm–3 of hydrochloric acid in a sample solution cations.
Then the integrated iodometric solid-phase spectrophotometry
Table 1 Influence of iodine concentration in the sample solution on method developed for the determination of ascorbic acid† was
parameters of calibration curve in the determination of ascorbic acid. For
parameter explanation see the text. tested on various juice samples. The results are collected in
Table 2 in comparison with the known reference indophenol
ciodine/
Regression equation
Linear range/
S2ad S2y titration method33 together with the significance of discrepancy
mol dm–3 mg dm–3 in the concentration obtained by the two methods. For three
5.0 × 10–5 A365 = 1.2 – 0.14casc 3.0–9.0 0.004 0.002 repetitive determinations of ascorbic acid following the proposed
7.5 × 10–5 A365 = 2.24 – 0.16casc 6.0–14.0 0.049 0.017 and the reference methods the statistical Student’s t- and F-tests
1.0 × 10–4 A365 = 3.1 – 0.17casc 8.0–16.0 0.022 0.014 revealed that there were no significant differences between the
1.5 × 10–4 A365 = 4.2 – 0.16casc 15.0–27.0 0.221 0.177
mean values and the variances for the two populations at the

Table 2 The results of ascorbic acid determination in real samples (n = 3, P = 0.95, Ftabl = 19, ttabl = 2.8).

casc as determined by solid-phase

casc as indicated on the casc as determined by titrimetry33
Juice spectrophotometry with PMM F t
package/mg dm–3
Found/mg dm–3 sr (%) Found/mg dm–3 sr (%)
Lemon pulp – 440 ± 50 4.6 450 ± 20 1.8 6.2 0.7
Orange no. 1 200 230 ± 30 5.3 210 ± 10 1.9 9.3 2.7
Orange no. 2 210 250 ± 30 4.8 260 ± 10 1.5 9.5 2.7
Carrot 300 340 ± 30 3.6 340 ± 10 1.2 9.0 0.4
Pineapple no. 1 800 930 ± 60 2.6 910 ± 60 2.6 1.0 1.8
Pineapple no. 2 800 980 ± 70 2.9 970 ± 70 2.9 1.0 1.6

† Procedure for determination of ascorbic acid. Ascorbic acid (0.05 g,
Dynemic Products, India) was dissolved in double distilled water
(50 cm3), the resulting 1 g dm–3 stock solution was consequently diluted
with distilled water to form working solutions, all the solutions were
prepared on the day of the experiment. The content of iodine ampoules
(Gosstandard, Russia) was diluted with distilled water in a 1000 cm3
measuring flask resulting in 0.025 mol dm–3 solution, the concentration
was measured by titration using the sodium thiosulfate standard solution.
Sodium 2,6-dichloroindophenolate (0.05 mg, Sigma-Aldrich) was
dissolved in hot distilled water (150 ml), cooled to room temperature,
diluted to 200 ml and filtered into an amber-colored bottle. Hydrochloric
acid (Sigma-Aldrich) was employed for the pH adjustment. All the
chemical reagents were used without purification. PMM plates were
placed into the test tubes with the solutions, the content was stirred for
5 min using a Multi Bio RS-24 programmable rotator (Biosan, Latvia) at
50 rpm. Then the plates were taken out and their absorption was measured
at 365 nm relative to the matrix source sample employing an Evolution
201 UV-VIS spectrophotometer (Thermo Fisher Scientific, USA). The
content of ascorbic acid was determined using the calibration curve built
under similar conditions.

– 137 –
 Mendeleev Commun., 2022, 32, 136–138
95% confidence level. Thus, the proposed method was considered
to be as accurate and precise as the reference one.
In summary, we demonstrated the feasibility of using PMM
for iodometric solid-phase spectrophotometric determination of
ascorbic acid in a 1.0–9.0 mg dm–3 concentration range with the
limit of detection 0.8 mg dm–3 as calculated by the 3s criterion.
The method suggested is easy to implement and requires only
standard spectrophotometric equipment. Compared to the
reference titration method, our approach offers the following
advantages: the rapidity and possibility of using it without any
preliminary sample processing for the intensely natural and
packaged pulpy juices.
This work was supported by the Tomsk Polytechnic University
SE Program.
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Received: 31st May 2021; Com. 21/6573